The Longevity Consortium continues to produce precedent-setting studies that seek to identify the factors that influence human longevity. Our validated experimental approach combines new multi-modal biochemical and analytical toolkits necessary to identify and characterize those factors, and produce new insights into how that knowledge can be leveraged to prevent disease, and enhance longevity. We will define coherent views of the major molecular pathways and processes causally influencing human longevity by using a combination of very large human cohorts of centenarians and healthy long-lived individuals, cross-species studies (designed to exploit evolutionary orthologous relationships), advanced molecular assays, chemoinformatic analyses, and systems data analytic methods. The Longevity Consortium Proteomic Project will identify proteomic signatures for longevity and aging by using recently developed deep proteomic profiling. We will apply our considerable expertise in whole proteome analyses to better understand the biology of longevity through use of mass spectrometric (MS) methods that enable precise protein measurements and improved computational proteomics, instrument performance and sample preparation, for robust quantification of a large fraction of endogenous proteins in serum or tissues. We have three aims: 1). We will use discovery proteomics by data-dependent analysis (DDA) and our innovative quantitative digital proteomics analysis (data-independent analysis (DIA or SWATH-MS)) of serum samples from human longevity cohorts, cells from long-lived and short-lived species of birds, primates, and rodents, as well as long-lived mutant mice and mice treated with drugs that extend longevity, to determine what longevity- associated proteomic signatures are present across experimental models. 2). We will employ extensive assessments of the post-translational modifications that are associated with longevity in human serum and mouse models. 3). We will use highly sensitive and specific targeted proteomics in MS instruments operated in selected reaction monitoring (SRM) mode combined with biomarker development to verify proteomic signatures of longevity that are identified in Aims 1 and 2, and through other Longevity Consortium projects and cores. To provide robust confidence in the elucidation of associations with longevity and related phenotypes, our statistical pipelines for processing and analyzing population proteomic data will include (a) rigorous methods to robustly estimate and test protein- and peptide-level associations, and (b) prioritization of candidate biomarkers. We will work in close collaboration with the other project and core components of the Longevity Consortium. Well characterized longevity associated proteins and/or post-translational modifications will be contributed to the Consortium for incorporation in systems-level analyses and ultimately for replication.